DE3930338A1 - METHOD FOR PRODUCING SORBIN ACID - Google Patents

Description

The present invention relates to a method for producing Sorbic acid, and particularly a process for the production of sorbic acid through microbiological oxidation of sorbic aldehyde.

Sorbic acid and its salts have preferably been used as food preservatives due to their excellent fungicidal activity. Industrially, sorbic acid has usually been prepared by reacting crotonaldehyde with a ketene to give a polyester via the β- lactone, which is an intermediate, and depositing the polyester by heat or an acid, or by using an ion exchange resin. However, the aforementioned process is not always advantageous in terms of production and economy, since the recovery and purification process of sorbic acid after the deposition of the polyester is complex, many steps and complex process control are required. Recently, to solve the aforementioned drawbacks, a method was disclosed in Japanese Unexamined Patent Publication No. 1 52 290/1988 in which sorbaldehyde was treated with a specific microorganism such as Pseudomonas to produce sorbic acid. However, when the process was carried out on an industrial scale, it was disadvantageous in terms of yield.

An object of the present invention is to provide a method for To provide production of sorbic acid in high yield.

These and other objects of the present invention will become apparent from the following description clarifies.

Serious efforts have been made to use sorbic acid a microbiological treatment.  

According to the present invention there is provided a method of manufacturing Sorbic acid provided the treatment of sorbic aldehyde with at least one microorganism selected from the group consisting of Mycobacterium, Rhodopseudomonas, Streptomyces, Acetobacter, Alcaligenes and Gluconobacter.

In the present invention, sorbic acid (2,4-hexadienoic acid) produced by microbiological oxidation of sorbic aldehyde (2,4- Hexadienal).

The microorganism used in the present invention is at least one microorganism selected from the groups consisting of Mycobacterium, Rhodopseudomonas, Streptomyces, Acetobacter, Alcaligenes and Gluconobacter.

Concrete examples of the microorganism are e.g. B.
Mycobacterium rhodochrous (Mycobacterium rhodochrous IFO 13161),
Rhodopseudomonas spheroides (Rhodopseudomonas spheroides IFO 12203),
Streptomyces albidoflavus (Streptomyces albidoflavus IFO 13010),
Acetobacter ascendens (Actobacter ascendens IFO 3188),
Acetobacter pasteurianus subsp lovanien (Acetobacter pasteurianus subsp lovanien IFO 13753),
Alcaligenes eutrophus (Alcaligenes eutrophus ATCC 17699),
Gluconobacter dioxyacetonics (Gluconobacter dioxyacetonics IFO 3271),
Gluconobacter gluconicus (Gluconobacter gluconicus IFO 3171),
Gluconobacter rubiginosus (Gluconobacter rubiginosus IFO 3244),
Gluconobacter suboxydans (Gluconobacter suboxydans IFO 3254, Gluconobacter suboxydans IFO 3256)
and similar. The expression "IFO" or "ATCC" indicates the depository "Institute for Fermentation, Osaka" or "American Type Culture Collection".

According to the present invention, any medium can be used for cultivation of the microorganism as mentioned above can be used as long as the medium contains a carbon source and a nitrogen source and the Microorganisms can grow in the medium.

Any compounds can be used as the carbon source, as long as these compounds promote the activity of the microorganism  Do not inhibit sorbic acid production. Examples of this Compounds used as a carbon source are e.g. B. Glucose, sucrose, ethanol, ethylene glycol, propylene glycol, 1,4-butanediol, Glycerin, acetaldehyde, acetic acid, propionic acid and the like.

As a nitrogen source z. B. meat extract, peptone, corn broth, Urea, ammonium sulfate, ammonium chloride, sodium nitrate and the like be used.

If necessary, inorganic salts such as phosphate, a magnesium salt, a potassium salt, an iron salt, a copper salt and a zinc salt or Nutrient substances necessary for washing the microorganism, be added to the medium.

If sorbinaldehyde is treated with the microorganism, one can Culture broth containing the grown microorganism in the medium, so can be used as it is, and also a suspension, in the microorganism, collected from the broth, suspended in water is physiological saline or a buffer solution is used will.

The concentration of sorbaldehyde in the reaction system is about 0.01 to about 10% by weight, preferably from about 0.05 to about 5% by weight,

Sorbaldehyde can be added to the broth or suspension in any way, e.g. B. in whole, in stages or continuously. For practical For reasons it is advantageous to add sorbaldehyde in portions. The Microorganism is used in an amount of about 0.5 to about 20 g (as Dry weight of the microorganism) used per liter of the suspension.

During the reaction of sorbaldehyde with the microorganism, this becomes Reaction system stirred under aerobic conditions. To achieve the aerobic conditions will be air or oxygen, and if necessary other gases, blown into the reaction system. The is preferably Oxygen content in the suspension not less than one ppm. Prefers  the reaction at a temperature of 10 to 70 ° C, especially preferably from 20 to 40 ° C over a period of about 0.1 to about 150 hours carried out. The pH of the system is preferably 4 to 9.

If necessary, a coenzyme such as PQQ (pyrroloquinolinequinone) or NAD (P) (nicotinamide adenine dinucleotide or its phosphate) surface active agent or an organic solvent Reaction system can be added.

In the present invention, the reaction can be done in any way be performed. For example, using either microorganisms that grow or those at rest or both can be done. Likewise, others can be immobilized Microorganisms or extracts from the microorganisms can be used.

After the end of the reaction, the reaction mixture is in the usual way cleaned to obtain the desired sorbic acid.

According to the present invention, sorbic acid can be obtained in high yield and high purity by oxidation of sorbaldehyde with the specific Microorganism can be produced.

The present invention will become more specific by the following examples described and explained. However, it goes without saying that the present invention is not limited to the examples and Various changes and modifications can be made without the Deviate the scope and content of the present invention.

Example 1

3 g meat extract, 10 g peptone, 5 g sodium chloride and 1 L water were added mixed to give a nutrient medium with a pH of 7. A Test tube was filled with 5 ml of the nutrient medium into the Mycobacterium rhodochrous IFO 13161 was inoculated in an amount of a platinum loop, and grown at 30 ° C for 24 hours with shaking, using a seed Culture broth received.  

A 500 ml Sakaguchi bottle was filled with 100 ml of the nutrient medium, in the 5 ml of the seed culture broth were inoculated, followed by cultivated with shaking at 30 ° C for 24 hours. After cultivation the cells were collected by centrifugation and washed with 0.1 molar Phosphate buffer (pH 7) washed, taking 200 mg (dry weight) cells received.

A 400 ml L-shaped tube was filled with 60 mg (dry weight) of the cells filled, whereupon 10 ml of 0.01 molar phosphate buffer were added and the Cells in the buffer were evenly suspended. Following were 10 mg of sorbaldehyde was added to the suspension to complete the reaction start. The reaction was carried out with shaking at 30 ° C 30 minutes. After that, the supernatant liquid removed from the reaction mixture by centrifugation and adjusted to pH of 2 adjusted with hydrochloric acid. The yield of sorbic acid obtained was measured by gas chromatography.

The results are shown in Table 1.

The conditions of gas chromatography were as follows:

Device:
HEWLETT PACKARD 5890
Quadrex bonded, sintered silica capillary column with methyl silicone 0.53 mm × 15 m × 8.0 µ film
Column temperature:
held at 80 ° C for eight minutes, raised to 210 ° C at a rate of 15 ° C / minute and held at 210 ° C for four minutes
Helium flow rate:
20 ml / minute
Dectection:
FID (flame ionization detector)

Examples 2 to 17

The procedure of Example 1 was repeated except that each Microorganism shown in table was used.  

The sorbic acid obtained was the same by means of gas chromatography Way is determined in Example 1. The results are in Table 1 shown.  

Table 1

In addition to the ingredients used in the examples, may include other ingredients in the examples, as previously in the Description will be explained, being essentially the same Results are achieved.

Suitable strains of the groups of the above-mentioned microorganisms for the method of the present invention are suitable can be via the the following procedures can be found:

3 g meat extract, 10 g peptone, 5 g sodium chloride and 1 l water mixed to obtain a nutrient medium with a pH of 7. A Test tube is filled with 5 ml of the nutrient medium into which the investigating microorganism inoculated in the amount of a platinum loop becomes. The mixture is then shaken at 30 ° C. for 24 hours cultivated to obtain a seed culture broth. A 500 ml Sakaguchi bottle is filled with 100 ml of the nutrient medium into which 5 ml of the Seed culture broth are inoculated, and is then shaken cultivated at 30 ° C for 24 hours. After cultivating, the Cells collected by centrifugation and with 10 ml 0.1 M phosphate buffer (pH 7), giving 200 mg (dry weight) of cells.

A 400 ml L-shaped tube is loaded with 60 mg (dry weight) of cells filled, to which 10 ml 0.01 molar phosphate buffer are added and the Cells are completely suspended in the buffer. Then 10 ml Sorbaldehyde was added to the suspension to start the reaction.

The reaction is carried out with shaking at 30 ° C. After 30 minutes the supernatant liquid is removed from the reaction mixture Centrifugation removed and adjusted to pH 2 with hydrochloric acid. The yield of the sorbic acid obtained is determined by gas chromatography measured.

If the amount of sorbaldehyde converted is 30% or more, may the microorganism is suitable in the process according to the invention be used. The microorganism is particularly suitable if the Yield is 40% or more, the is particularly suitable Microorganism if the yield is about 50% or more.

Claims (2)

1. A process for the preparation of sorbic acid, characterized in that sorbaldehyde is treated with at least one microorganism selected from the group consisting of Mycobacterium, Rhodopseudomonas, Streptomyces, Acetobacter, Alcaligenes and Gluconobacter. 2. The method according to claim 1, characterized in that this Microorganism is at least one microorganism selected from the Group consisting of Mycobacterium rhodochrous, Rhodopseudomonas spheroides, Streptomyces albidoflavus, Acetobacter ascendens, Acetobacter pasteurianus subsp lovanien, Alcaligenes eutrophus, Gluconobacter dioxyacetonics, Gluconobacter gluconicus, Gluconobacter oxydance, Gluconobacter rubiginosus and Gluconobacter suboxydans.